The long term goals of this research are to understand the structure, intracellular transport and metabolism and function of the major DNA binding protein of herpes simplex virus (HSV). In this proposal we outline our approaches to determine whether there are distinct functional domains of the protein involved in the transport of the protein to the cell nucleus as opposed to its binding to viral DNA. We will use genetic and biochemical analysis of the viral gene product in these experiments. We also propose to further map and define the gene encoding this protein through the mapping and analysis of its mRNA transcript. We also propose to use this system as a model for the study of the nuclear transport of proteins. We will define the cellular interactions of wild type and mutant viral proteins with the host cell to understand the route and mechanism of accumulation of specific proteins in the cell nucleus. This viral gene product may also be of clinical importance in that an antigenically related protein has been detected in HSV transformed cells, cervical carcinoma tissue and vulvar carcinoma tissue. Thus this viral gene product may play a role in the transformation or tumorigenic process. Whether or not this protein is involved in the initiation of these neoplastic converstions, the protein could affect the metabolism of the transformed or tumor cells. We propose experiments to study the properties of the viral gene product to better predict its effect on the metabolism of a transformed cell.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA026345-07
Application #
3167261
Study Section
Virology Study Section (VR)
Project Start
1979-07-01
Project End
1987-12-31
Budget Start
1986-01-01
Budget End
1986-12-31
Support Year
7
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Harvard University
Department
Type
Schools of Medicine
DUNS #
082359691
City
Boston
State
MA
Country
United States
Zip Code
02115
Uprichard, Susan L; Knipe, David M (2003) Conformational changes in the herpes simplex virus ICP8 DNA-binding protein coincident with assembly in viral replication structures. J Virol 77:7467-76
Taylor, Travis J; McNamee, Elizabeth E; Day, Cheryl et al. (2003) Herpes simplex virus replication compartments can form by coalescence of smaller compartments. Virology 309:232-47
Taylor, Travis J; Knipe, David M (2003) C-terminal region of herpes simplex virus ICP8 protein needed for intranuclear localization. Virology 309:219-31
Brockman, Mark A; Knipe, David M (2002) Herpes simplex virus vectors elicit durable immune responses in the presence of preexisting host immunity. J Virol 76:3678-87
Zhou, Changhong; Knipe, David M (2002) Association of herpes simplex virus type 1 ICP8 and ICP27 proteins with cellular RNA polymerase II holoenzyme. J Virol 76:5893-904
Da Costa, X J; Morrison, L A; Knipe, D M (2001) Comparison of different forms of herpes simplex replication-defective mutant viruses as vaccines in a mouse model of HSV-2 genital infection. Virology 288:256-63
McNamee, E E; Taylor, T J; Knipe, D M (2000) A dominant-negative herpesvirus protein inhibits intranuclear targeting of viral proteins: effects on DNA replication and late gene expression. J Virol 74:10122-31
Da Costa, X; Kramer, M F; Zhu, J et al. (2000) Construction, phenotypic analysis, and immunogenicity of a UL5/UL29 double deletion mutant of herpes simplex virus 2. J Virol 74:7963-71
de Bruyn Kops, A; Uprichard, S L; Chen, M et al. (1998) Comparison of the intranuclear distributions of herpes simplex virus proteins involved in various viral functions. Virology 252:162-78
Uprichard, S L; Knipe, D M (1997) Assembly of herpes simplex virus replication proteins at two distinct intranuclear sites. Virology 229:113-25

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